European journal of protistology最新文献

Acanthamoeba castellanii is a free-living amoeba and an opportunistic pathogen for humans that can cause encephalitis and, more commonly, Acanthamoeba keratitis. During its life cycle, A. castellanii may present as proliferative and infective trophozoites or resistant cysts. The adhesion of trophozoites to host cells is a key first step in the pathogenesis of infection. A major virulence protein of Acanthamoeba is a mannose-binding protein (MBP) that mediates the adhesion of amoebae to cell surfaces. Ectophosphatases are ecto-enzymes that can dephosphorylate extracellular substrates and have already been described in several microorganisms. Regarding their physiological roles, there is consistent evidence that ectophosphatase activities play an important role in parasite-host interactions. In the present work, we identified and biochemically characterized the ectophosphatase activity of A. castellanii. The ectophosphatase activity is acidic, stimulated by magnesium, cobalt and nickel, and presents the following apparent kinetic parameters: K_m = 2.12 ± 0.54 mM p-NPP and V_max = 26.12 ± 2.53 nmol p-NP × h⁻¹ × 10^-6 cells. We observed that sodium orthovanadate, ammonium molybdate, sodium fluoride, and inorganic phosphate are able to inhibit ectophosphatase activity. Comparing the two stages of the A. castellanii lifecycle, ectophosphatase activity is significantly higher in trophozoites than in cysts. The ectophosphatase activity is stimulated by mannose residues and is significantly increased when trophozoites interact with LLC-MK2 cells. The inhibition of ectophosphatase by pretreatment with sodium orthovanadate also inhibits the adhesion of trophozoites to epithelial cells. These results allow us to conclude that the ectophosphatase activity of A. castellanii is somehow important for the adhesion of trophozoites to their host cells. According to our data, we believe that the activation of MBP by mannose residues triggers the stimulation of ectophosphatase activity to facilitate the adhesion process.

Apocarchesium rosettum and A. arndti were originally discovered in Japan (Lake Biwa) and Germany (River Rhine), respectively. We report the first record of A. arndti in the Danube and provide a detailed description of its colony development. Our findings support the theory of moderate endemicity and reveal a new, smaller zooid type in A. arndti. This zooid remains attached to the colony, connected to the stalk myoneme but lacks an aboral ciliary wreath. Unlike microzooids, it is incapable of leaving the colony. It exhibits a less spherical shape and arises from the fourth division of the colony-founder cell. Although its specific function is unknown, it is hypothesized to support the stalk dish. Our results have significant implications for understanding the systematics of vorticellids, suggesting their ancestral nature as colonial organisms characterized by a helically contracting stalk myoneme. Furthermore, the exclusive retention of the stalk myoneme by the parental cell after binary fission may serve as a synapomorphy for the Vorticellidae. We provide a descriptive analysis of the ecological environment and microhabitat of A. arndti in the Danube, revealing its preference for well-developed, detritus-rich biofilms during summer, absence in late winter and spring, emergence during peak summer, and subsequent decline until mid-winter.

Arcellinida is ascending in importance in protistology, but description of their diversity still presents multiple challenges. Furthermore, applicable tools for surveillance of these organisms are still in developing stages. Importantly, a good database that sets a correspondence between molecular barcodes and species morphology is lacking. Cytochrome oxidase (COI) has been suggested as the most relevant marker for species discrimination in Arcellinida. However, some major groups of Arcellinida are still lacking a COI sequence. Here we expand the database of COI marker sequences for Arcellinids, using single-cell PCR, transcriptomics, and database scavenging. In the present work, we added 24 new Arcellinida COI sequences to the database, covering all unsampled infra- and suborders. Additionally, we added six new SSUrRNA sequences and described four new species using morphological, morphometrical, and molecular evidence: Heleopera steppica, Centropyxis blatta, Arcella uspiensis, and Cylindrifflugia periurbana. This new database will provide a new starting point to address new research questions from shell evolution, biogeography, and systematics of arcellinids.

Acanthamoeba castellanii is a free-living amoeba that acts as an opportunistic pathogen for humans and is the pathogenic agent of Acanthamoeba keratitis (AK). A. castellanii may present as proliferative and infective trophozoites or as resistant cysts during their life cycle. The immune response against AK is still poorly explored; however, it is well established that macrophages and neutrophils play essential roles in controlling corneal infection during the disease outcome. The release of NETs is one of the innate immune strategies to prevent parasite infection, especially when neutrophils interact with microorganisms that are too large to be phagocytosed, which is the case for amoeba species. The present work demonstrated that A. castellanii trophozoites can trigger NET formation upon in vitro interaction with neutrophils. Using DNase as a control, we observed increased parasite survival after coinciding with neutrophils, which may be correlated with NET degradation. Indeed, A. castellanii trophozoites degrade the NET DNA scaffold. Molecular analysis confirmed the occurrence of a 3′-nucleotidase/nuclease (3′-NT/NU) in the A. castellanii genome. We also demonstrated that trophozoites exhibit significantly higher 3′-NT/NU activity than cysts, which cannot trigger NET release. Considering that previous studies indicated the pathological role of 3′-NT-/NU in parasite infection, we suggest that this enzyme may act as the mechanism of escape of A. castellanii trophozoites from NETs.

Trachelocercids are common ciliates in marine sandy intertidal zones, however, only few species have been described using modern taxonomic techniques, with several earlier descriptions still lacking primary data, such as a detailed description of the oral ciliature. For this reason, systematics of the family Trachelocercidae Kent, 1881 resulted in paraphyletic genera and needs a critical revision. As an attempt to fill the gap of knowledge for this group, this study presents one new species description, Tracheloraphis apodiscolor n. sp., along with redescriptions of three lesser-known trachelocercid species, Trachelocerca bodiani (Dragesco, 1963) Dragesco, 2002, Trachelocerca ditis (Wright, 1982) Foissner & Dragesco, 1996 and Prototrachelocerca fasciolata (Sauerbrey, 1928) Foissner, 1996, collected from coastal marine habitats at Qingdao (China), using live and protargol-stained specimens. Tracheloraphis apodiscolor n. sp. can be distinguished from its closest related congener, T. discolor, by a shorter body length and fewer somatic kineties. Molecular data of T. apodiscolor n. sp. and T. bodiani were provided for the first time. Additionally, based on the Qingdao populations, we provide improved diagnoses for T. ditis and P. fasciolata. Phylogenetic analyses were performed and discussed, as there is strong support for paraphyly of the genera Tracheloraphis and Trachelocerca. In general, more integrative studies based on taxonomy, ultrastructure as well as molecular data are needed to properly shed light on the systematics of trachelocercids.

Diplomonadida is a lineage of anaerobic protists belonging to Fornicata, Metamonada. Most diplomonads are endobiotic or parasitic, such as Giardia intestinalis, which is a famous human pathogen, but several free-living species exist as well. Although it has been proposed that the free-living diplomonads are descendants of endobiotic organisms and thus interesting from the evolutionary point of view, they have been largely neglected. We obtained 58 cultures of free-living diplomonads belonging to four genera (Hexamita, Trepomonas, Gyromonas, and Trimitus) and six strains of endobiotic diplomonads and analyzed their SSU rRNA gene sequences. We also studied light-microscopic morphology of selected strains and the ultrastructure of Trepomonas rotans for the first time. Our phylogenetic analysis showed that the genus Hexamita, and, possibly, also the genus Trepomonas, are polyphyletic. Trepomonas rotans, which may represent a novel genus, is unique among Diplomonadida by having the cell covered in scales. Our results suggest that the evolution of the endobiotic life style and cell organization in diplomonads is more complicated than previously thought.

Although free-living ciliated protists (Ciliophora) commonly inhabit aquatic and terrestrial biotopes of mild to moderate salinity (PSS-practical salinity scale S = 10–150), very few have adapted to life at more extreme salinities (>150 to saturation). Such extreme halophiles or halotrophs are of interest from the standpoints of evolution, cell physiology, ecology, and even astrobiology. In this work, we present the morphology, 18S rRNA gene sequence, and phylogenetic analysis of a novel spathidiid ciliate (Ciliophora, Litosatomatea) that thrives in saturated brines of Maharloo Lake, Iran. Based on its unique combination of morphologic features, its molecular characterization, and its unusual ecology, it is assigned to a newly erected monotypic genus incertae sedis in order Spathidiida. The new species differs from other spathidiids by: a cell with a very short acutely pointed tail, the absence of a circumoral kinety with, instead, inclined, interrupted circumoral kinetofragments at the anterior end of somatic kineties and brush rows abutting the oral bulge, the absence of a contractile vacuole, a heteromorphic dorsal brush comprising five or six rows, and an extreme (at or near saturation) hypersaline habitat. We briefly discuss the persistent uncertainties regarding the phylogenetic relationships within the order Spathidiida.

Coccidiosis, a serious intestinal parasitic disease caused by Eimeria spp., can result in huge annual economic losses to the poultry industry worldwide. At present, coccidiosis is mainly controlled by anticoccidial drugs. However, drug resistance has developed in Eimeria because of the long-term and unreasonable use of the drugs currently available. In our previous study, RNA-seq showed that the expression of methionine aminopeptidase1 (EtMetAP1) was up-regulated in diclazuril-resistant (DZR) and maduramicin-resistant (MRR) strains compared to drug-sensitive (DS) strain of Eimeria tenella. In this study, EtMetAP1 was cloned and expressed, and the function and characteristics of the EtMetAP1 protein were analyzed. The transcription and translation levels of EtMetAP1 in DS strain of E. tenella at different developmental stages were analyzed by qPCR and western blotting. We found that the transcription and translation levels of EtMetAP1 in second-generation merozoites (SM) were higher than those of the other three stages (unsporulated oocyst, sporulated oocyst, and sporozoites). Simultaneously, qPCR was used to analyze the mRNA transcription levels of EtMetAP1 in DS, DZR, MRR, and salinomycin-resistant (SMR) strain. The results showed that compared to the sensitive strain, the transcription levels of EtMetAP1 in DZR and MRR were up-regulated. There was no significant difference in transcription level in SMR. Indirect immunofluorescence localization showed that the protein was mainly localised in the cell membrane and cytoplasm of sporozoites and SM. An invasion inhibition test showed that anti-rEtMetAP1 polyclonal antibody could effectively inhibit the sporozoite invasion of host cells. These results suggest that the protein may be involved in the growth and development of parasites in host cells, the generation of drug resistance, and host cell invasion.

Maintaining high levels of biological diversity in various ecosystems is necessary for stable functioning of the Earth's biosphere. The article describes diversity and ecology of heterotrophic siliceous protists – rotosphaerids, colourless free-living thaumatomonad flagellates, and centrohelid heliozoans – in Arctic waters located of Asian Russia. Samples were collected in the mouths of the Olenyok, Yana, Indigirka, Kolyma Rivers – and small freshwater ponds near Settlement of Tiksi in Yakutia. Based on electron microscopy, 35 centrohelids (Heliozoa), 11 rotosphaerid species as well as four thaumatomonad flagellate species were found in the region. Seven species were recorded in Russia for the first time: Rabdiaster multicosta, Rabdiaster reticulata, Turriplaca denticulata, Choanocystis cf. cordiformis parvula, Raineriophrys pteromorphos, Pseudoraphidocystis glutinosa and Pseudoraphidiophrys formosa. For 43 species, the study area is the most northern location they were described. Morphological details of scales are discussed for selected species, particularly for widespread species of rotosphaerids – Pinaciophora fluviatilis. The literature on hydrochemical conditions was analyzed to find taxa with high sensitivity towards environmental changes. Such species could be further used for monitoring plankton, recovering evolutionary changes, and reconstructing past environments.